Saliva is an ideal medium in which to measure cortisol in children. However, there are very few data reporting salivary cortisol or cortisone concentrations in healthy children since the introduction of liquid chromatography-mass spectrometry (LC-MS/MS) to routine laboratory practice. Early morning serum cortisol, salivary cortisol and cortisone were measured on fasting samples, and salivary hormones were measured in samples collected every 2hours during waking hours, and 30minutes after waking the following morning. 43 healthy paediatric volunteers (19 female), median age 11.5years, range 6.2-18.7, participated. Early morning serum cortisol (265nmol/L, 156-516) correlated strongly with salivary cortisol (4.7nmol/L, 1.1-14.6) and cortisone (28.8nmol/L, 11.7-56.6), P<.0001 for both. Serum cortisol, salivary cortisol and salivary cortisone correlated directly with age (P<.0001, P=.002 and P=.015, respectively), and salivary cortisone/cortisol ratio correlated indirectly with age (P=.007). Between 08.00 and 21.00, area under the curve for salivary cortisol (mean±1 SD) was 41.8±19.1 and for cortisone 213.0±61.2. Salivary cortisol was undetectable in 25/130 (19%) of samples collected after 13.00, while cortisone was always detectable. Salivary cortisol and cortisone concentrations are strongly related to serum cortisol concentrations; however, cortisone may be a preferable measure as cortisol is often undetectable. Age may be an important factor in the interpretation of early morning cortisol measurements made in serum and saliva.